Novel methods for β-secretase kinetic and inhibition assays with unlabelled substrates using capillary electrophoresis with various detection techniques
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| Year of publication | 2016 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | In our work we present novel method for ß-secretase kinetic and inhibition assays using unlabelled decapeptide, which contains so called Swedish mutation of amyloid precursor protein, as substrate using capillary electrophoresis coupled with mass spectrometry. Thanks to low sample consumption and high sensitivity the enzyme reaction can be carried out in as small volume as only 100 µl in just 30 min. These features together with deploying unlabelled substrate ensure low cost per analysis. What is more, our results show high difference between kinetic parameters of unlabelled substrate in comparison with kinetic parameters of fluorescently labelled substrates published in literature by two orders of magnitude even though the incubation conditions used were comparable. On the other hand determined inhibition parameters of two reported inhibitors show excellent agreement with literature data. These results were confirmed by comprehensive kinetic study of BACE1 carried out using different detection techniques. |
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